Considering the cytotoxic properties of Schiff base ligands, we re-synthesized a gold(III) complex with a tetradentate Schiff base ligand and investigated its cytotoxic effects using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method and sulforhodamine B staining method. The results showed that the gold(III) Schiff base complex can inhibit the growth of Chinese hamster ovary (CHO) cancer cell lines. However, unlike cisplatin, it does not induce cell death in NIH 3T3 normal cells. Real-time polymerase chain reaction (PCR) results revealed that the gold(III) Schiff base complex increased the Bak1/Bclx ratio in CHO cell line by 3.53-fold compared to the control, while cisplatin increased the anti-apoptotic Bclx gene by 31.8 times and the treatment resistance AKT1 gene by 430.8 times the control. However, cisplatin only activated the extrinsic pathway of apoptosis through the caspase-3 gene. The expression levels of TLR6 and TNF-α genes in normal cells showed that both the gold(III) Schiff base complex and cisplatin did not cause inflammation in NIH 3T3 cell lines. All the results indicate that the new complex could be a promising candidate to replace cisplatin. It has been shown that it induces apoptosis and prevents the development of resistance in cancer cells. Additionally, it does not cause inflammation in normal cells. ligands resynthesized re synthesized goldIII gold III gold(III 34,5dimethylthiazol2yl2,5diphenyltetrazolium 345dimethylthiazol2yl25diphenyltetrazolium dimethylthiazolyldiphenyltetrazolium 3 4,5 dimethylthiazol 2 yl 2,5 diphenyltetrazolium 4 5 bromide (CHO lines However T 3T Realtime Real time PCR (PCR Bak1Bclx BakBclx Bak1 Bak 3.53fold 353fold fold 3.53 53 control antiapoptotic anti apoptotic 318 31 8 31. AKT 4308 430 430. caspase3 caspase caspase- TLR TNFα TNF α Additionally 34 5dimethylthiazol2yl2 5diphenyltetrazolium 45 4, 25 2, 53fold 353 3.5 43 dimethylthiazolyl 5dimethylthiazol2yl 35 3.

RESUMO Objetivo: Verificar a utilização e identificar as vantagens dos métodos moleculares para diagnóstico de infecções congênitas no líquido cefalorraquidiano de neonatos. Fontes de dados: A revisão foi registrada na base PROSPERO (International Prospective Register of Systematic Reviews) sob CRD42021274210. A busca bibliográfica foi realizada nas bases de dados PubMed, Biblioteca Virtual em Saúde/ Centro Latino-Americano e do Caribe de Informação em Ciências da Saúde (BVS/BIREME), Scopus, Web of Science, Excerpta Medica database (EMBASE), Cochrane, ProQuest, e EBSCOhost. A busca foi feita no período de agosto a outubro de 2021 e atualizada em dezembro de 2022, respeitando as orientações do Preferred Reporting Items for Systematic Reviews e Meta-Analyises (PRISMA). A sequência da seleção dos estudos foi: 1) Remoção de duplicatas; 2) Exame de títulos e resumos; 3) Recuperação dos textos completos potencialmente relevantes; e 4) Avaliação do texto completo conforme critérios de elegibilidade por dois autores independentes. O critério de inclusão considerou ensaios clínicos randomizados e não randomizados, estudos longitudinais, transversais, revisados por pares, estudos em humanos, publicados em inglês, espanhol, italiano e português, com recém-nascidos de até 28 dias que sofreram neuroinfecções congênitas pelos agentes toxoplasmose, rubéola, citomegalovírus, herpes simples (TORCH), e outros como Treponema pallidum, Zika, parvovírus B-19, varicela zoster, Epstein-Barr, e SARS-CoV-2, diagnosticadas por reação em cadeia de polimerase (PCR). Dois avaliadores extraíram as seguintes informações: autor, ano de publicação, nacionalidade, sujeitos, tipo de estudo, métodos, resultados e conclusão. Síntese dos dados: O patógeno mais estudado foi Herpes Simples. Muitos artigos relataram somente sintomas iniciais inespecíficos, motivando a coleta de líquido cefalorraquidiano e realização da PCR para investigação etiológica. Conclusões: Os métodos moleculares são eficazes para detectar o genoma do patógeno no líquido cefalorraquidiano, o que pode impactar na evolução clínica e no prognóstico neurológico. Objetivo neonatos International CRD42021274210 CRD PubMed LatinoAmericano Latino Americano BVS/BIREME, BVSBIREME BVS/BIREME , BVS BIREME (BVS/BIREME) Scopus Science EMBASE, EMBASE (EMBASE) Cochrane ProQuest EBSCOhost 202 2022 MetaAnalyises Meta Analyises PRISMA. PRISMA . (PRISMA) 1 duplicatas 2 resumos 3 relevantes 4 independentes longitudinais transversais pares humanos inglês espanhol português recémnascidos recém nascidos toxoplasmose rubéola citomegalovírus TORCH, TORCH (TORCH) pallidum Zika B19, B19 B 19, 19 B-19 zoster EpsteinBarr, EpsteinBarr Epstein Barr, Barr Epstein-Barr SARSCoV2, SARSCoV2 SARSCoV SARS CoV 2, SARS-CoV-2 PCR. (PCR) informações autor publicação nacionalidade sujeitos estudo conclusão Simples inespecíficos etiológica Conclusões neurológico CRD4202127421 (BVS/BIREME (EMBASE 20 (PRISMA (TORCH B1 B-1 SARS-CoV- (PCR CRD420212742 B- SARS-CoV CRD42021274 CRD4202127 CRD420212 CRD42021 CRD4202 CRD420 CRD42 CRD4

ABSTRACT Objective: To verify the use and identify advantages of molecular methods for congenital infections diagnosis in cerebrospinal fluid of neonates. Data source: The review was registered in the International Prospective Register of Systematic Reviews (PROSPERO), under CRD42021274210. The literature search was performed in databases: PubMed, Virtual Health Library/ Latin American and Caribbean Center on Health Sciences Information (VHL/BIREME), Scopus, Web of Science, Excerpta Medica database (EMBASE), Cochrane, ProQuest, and EBSCOhost. The search was carried out from August to October 2021 and updated in December 2022, respecting the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The selection sequence was: 1) Duplicate title removal; 2) Examination of titles and abstracts; 3) Full-text retrieval of potentially relevant reports; and 4) Evaluation of the full text according to eligibility criteria by two independent authors. Inclusion criteria considered randomized and non-randomized control trials, longitudinal, cross-sectional, and peer-reviewed studies in humans, published in English, Spanish, Italian, and Portuguese, with newborns up to 28 days old who had congenital neuroinfections by toxoplasmosis, rubella, cytomegalovirus, herpes simplex (TORCH), and others such as Treponema pallidum, Zika, parvovirus B-19, varicella zoster, Epstein-Barr, and SARS-CoV2, diagnosed by polymerase chain reaction (PCR). Two evaluators extracted the following information: author, year of publication, nationality, subjects, study type, methods, results, and conclusion. Data synthesis: The most studied pathogen was herpes simplex. Several articles reported only nonspecific initial symptoms, motivating the collection of cerebrospinal fluid and performing PCR for etiological investigation. Conclusions: Molecular methods are effective to detect pathogen genomes in cerebrospinal fluid, which can impact clinical evolution and neurological prognosis. Objective neonates source PROSPERO, PROSPERO , (PROSPERO) CRD42021274210 CRD databases PubMed Library VHL/BIREME, VHLBIREME VHL/BIREME VHL BIREME (VHL/BIREME) Scopus Science EMBASE, EMBASE (EMBASE) Cochrane ProQuest EBSCOhost 202 2022 MetaAnalyses Meta Analyses PRISMA (PRISMA guidelines 1 removal 2 abstracts 3 Fulltext Full reports 4 authors nonrandomized non trials longitudinal crosssectional, crosssectional cross sectional, sectional cross-sectional peerreviewed peer reviewed humans English Spanish Italian Portuguese toxoplasmosis rubella cytomegalovirus TORCH, TORCH (TORCH) pallidum Zika B19, B19 B 19, 19 B-19 zoster EpsteinBarr, EpsteinBarr Epstein Barr, Barr Epstein-Barr SARSCoV2, SARSCoV2 SARSCoV SARS CoV2, CoV2 CoV SARS-CoV2 PCR. . (PCR) information author publication nationality subjects type results conclusion synthesis symptoms investigation Conclusions prognosis (PROSPERO CRD4202127421 (VHL/BIREME (EMBASE 20 (TORCH B1 B-1 SARS-CoV (PCR CRD420212742 B- CRD42021274 CRD4202127 CRD420212 CRD42021 CRD4202 CRD420 CRD42 CRD4

Abstract Introduction: This study investigated variables associated with mortality in kidney transplant recipients (KTRs) diagnosed with post-transplant lymphoproliferative disease (PTLD) and a simultaneous Epstein-Barr virus (EBV) viremia. Methods: This was a retrospective cohort study enrolling KTRs diagnosed with PTLD between 2018 and 2020. Outcome: death within two years after diagnosis. Results: Among 1,625 KTRs who collected EBV viremia (by PCR, 2018–2020) for any reason, 238 (14.6%) had a positive viral load and 41 (17.2%) simultaneous PTLD. These 41 patients were 40.1 years old at diagnosis and 8.6 years after transplantation; 26.8% were induced with rATG and 92.7% were maintained on tacrolimus and azathioprine (TAC/AZA) as immunosuppressive regimen. Lymph nodes (75.6%) was the most common site of PTLD, followed by the gastrointestinal tract (48.8%), with 61.0% at Lugano stage IV and 80.5% monomorphic PTLD. The mean EBV viral load was 12,198 IU/mL. One- and two-year patient survival post-diagnosis was 60.4% and 46.8%, respectively. In the Cox regression analysis, age at PTLD diagnosis (HR for each year = 1.039; p < 0.001) and EBV viral load (HR for each log = 1.695; p = 0.026) were associated with risk of death. Conclusion: This study suggests that in patients predominantly on TAC/AZA, PTLD with simultaneous EBV positive viral load is a late event, and worse survival is associated with older age and EBV viral load at diagnosis. Introduction (KTRs posttransplant post (PTLD EpsteinBarr Epstein Barr (EBV Methods 201 2020 Outcome Results 1625 1 625 1,62 PCR 2018–2020 20182020 reason 23 14.6% 146 14 6 (14.6% 4 17.2% 172 17 2 (17.2% 401 40 40. 86 8 8. transplantation 268 26 26.8 927 92 7 92.7 TAC/AZA TACAZA TAC AZA (TAC/AZA regimen 75.6% 756 75 (75.6% 48.8%, 488 48.8% , 48 (48.8%) 610 61 0 61.0 805 80 5 80.5 12198 12 198 12,19 IUmL IU mL IU/mL One twoyear postdiagnosis 604 60 60.4 468 46 46.8% respectively analysis HR 1.039 1039 039 0.001 0001 001 1.695 1695 695 0.026 0026 026 Conclusion event 20 202 162 62 1,6 2018–202 2018202 14.6 (14.6 17.2 (17.2 26. 9 92. 75.6 (75.6 48.8 (48.8% 61. 80. 1219 19 12,1 60. 46.8 1.03 103 03 0.00 000 00 1.69 169 69 0.02 002 02 16 1, 2018–20 201820 14. (14. 17. (17. 75. (75. 48. (48.8 121 12, 46. 1.0 10 0.0 1.6 2018–2 20182 (14 (17 (75 (48. 1. 0. 2018– (1 (7 (48 ( (4

Resumo Introdução: Este estudo investigou variáveis associadas à mortalidade em receptores de transplante renal (RTR) diagnosticados com doença linfoproliferativa pós-transplante (PTLD, do inglês post-transplant lymphoproliferative disease) e viremia simultânea pelo vírus Epstein-Barr (EBV). Métodos: Estudo de coorte retrospectivo incluindo RTR diagnosticados com PTLD entre 2018 e 2020. Desfecho: óbito em até dois anos após diagnóstico. Resultados: Entre 1.625 RTR que realizaram coleta de viremia para EBV (por PCR, 2018–2020) por qualquer motivo, 238 (14,6%) apresentaram carga viral positiva e 41 (17,2%) PTLD simultânea. Esses 41 pacientes tinham em média 40,1 anos ao diagnóstico e 8,6 anos após o transplante; 26,8% foram induzidos com rATG e 92,7% foram mantidos com tacrolimus e azatioprina (TAC/AZA) como regime imunossupressor. Linfonodos (75,6%) foram o local mais comum de PTLD, seguidos pelo trato gastrointestinal (48,8%), com 61,0% no estágio IV de Lugano e 80,5% PTLD monomórfica. A carga viral média do EBV foi 12.198 UI/mL. A sobrevida dos pacientes em um e dois anos após o diagnóstico foi 60,4% e 46,8%, respectivamente. Na análise de regressão de Cox, a idade ao diagnóstico de PTLD (HR para cada ano = 1,039; p < 0,001) e a carga viral do EBV (HR para cada log = 1,695; p = 0,026) foram associadas ao risco de óbito. Conclusão: Este estudo sugere que, em pacientes predominantemente em uso de TAC/AZA, a PTLD com carga viral simultânea positiva para EBV é um evento tardio, e a pior sobrevida está associada à idade mais avançada e à carga viral de EBV no momento do diagnóstico. Introdução (RTR póstransplante pós (PTLD posttransplant post transplant disease EpsteinBarr Epstein Barr EBV. . (EBV) Métodos 201 2020 Desfecho Resultados 1625 1 625 1.62 PCR 2018–2020 20182020 motivo 23 14,6% 146 14 6 (14,6% 4 17,2% 172 17 2 (17,2% 401 40 40, 86 8 8, 268 26 26,8 927 92 7 92,7 TAC/AZA TACAZA TAC AZA (TAC/AZA imunossupressor 75,6% 756 75 (75,6% 48,8%, 488 48,8% , 48 (48,8%) 610 61 0 61,0 805 80 5 80,5 monomórfica 12198 12 198 12.19 UImL UI mL UI/mL 604 60 60,4 468 46 46,8% respectivamente Cox HR 1,039 1039 039 0,001 0001 001 1,695 1695 695 0,026 0026 026 Conclusão tardio (EBV 20 202 162 62 1.6 2018–202 2018202 14,6 (14,6 17,2 (17,2 26, 9 92, 75,6 (75,6 48,8 (48,8% 61, 80, 1219 19 12.1 60, 46,8 1,03 103 03 0,00 000 00 1,69 169 69 0,02 002 02 16 1. 2018–20 201820 14, (14, 17, (17, 75, (75, 48, (48,8 121 12. 46, 1,0 10 0,0 1,6 2018–2 20182 (14 (17 (75 (48, 1, 0, 2018– (1 (7 (48 ( (4

Resumen (Objetivo) Tryapanosoma cruzi, un agente causal de la enfermedad de Chagas, es un parásito que presenta una alternancia en su ciclo de vida entre un hospedador invertebrado (triatomino) y uno vertebrado (mamífero). Varios estudios han demostrado que en T. cruzi, HSP83 (homólogo de HSP90) es esencial para la división celular y el control de la respuesta al estrés térmico. Esta investigación se concentró en el estudio de la clonación, la caracterización bioinformática y la expresión del gen de la proteína de choque térmico HSP83 de T. cruzi para estudios posteriores sobre señalización. (Metodología) El ARN fue extraído de epimagostigotes T. cruci (clon EPm6/MOHM/VE/2007/ 6c), utilizando un kit comercial. El ADNc que codifica HSP83 se obtuvo utilizando RT-PCR, a partir del ARNm extraído, para lo cual se diseñaron los cebadores con base en la secuencia HSP83 de la cepa T. cruzi CL Brener. La clonación se realizó usando pGEM®T-Easy y se subclonó en el vector de expresión pQE30. Se efectuó la caracterización bioinformática y de secuencias. El gen se expresó y la proteína recombinante se purificó por cromatografía de afinidad, igual que se identificó por inmunotransferencia. (Resultados) El análisis de secuencia mostró similitud con el gen HSP83 de Trypanosoma cruzi y se observaron dominios HSP, así como epítopos B en la secuencia. Después de 3 horas de inducción con IPTG, se obtuvo una proteína recombinante con un peso aproximado de 83 kDa. La reacción de inmunotransferencia con suero hiperinmune anti-epimastigote de T. cruzi permitió la detección de una sola banda con un peso molecular de aproximadamente 83 kDa. (Conclusiones) Todos los resultados indican que se logró la clonación y caracterización de HSP83 de Trypanosoma cruzi.

Resumo (Objetivo) O Tryapanosoma cruzi, agente causador da doença de Chagas, é um parasita que apresenta uma alternância em seu ciclo de vida entre um hospedeiro invertebrado triatomíneo) e um vertebrado(mamífero). Vários estudos demonstraram que, no T. cruzi, a HSP83 (homóloga da HSP90) é essencial para a divisão celular e o controle da resposta ao estresse térmico. Esta pesquisa concentrou-se no estudo da clonagem, caracterização bioinformática e expressão do gene da proteína de choque térmico HSP83 do T. cruzi para estudos adicionais de sinalização. (Metodologia) O RNA foi extraído de epimastigotas de T. cruzi (clone EPm6/MOHM/VE/2007/ 6c) usando um kit comercial. O cDNA que codifica a HSP83 foi obtido por meio de RT-PCR a partir do mRNA extraído, para o qual os primers foram projetados com base na sequência da HSP83 da cepa T. cruzi CL Brener. A clonagem foi realizada com o pGEM®T-Easy e subclonada no vetor de expressão pQE30. Foi realizada a caracterização bioinformática e de sequência. O gene foi expresso e a proteína recombinante foi purificada por cromatografia de afinidade e identificada por imunotransferência. (Resultados) A análise da sequência mostrou similaridade com o gene HSP83 do Trypanosoma cruzi e os domínios HSP, bem como os epítopos B, foram observados na sequência. Após 3 horas de indução com IPTG, foi obtida uma proteína recombinante com peso aproximado de 83 kDa. A imunotransferência com soro hiperimune antiepimastigota de T. cruzi permitiu a detecção de uma única banda com peso molecular de aproximadamente 83 kDa. (Conclusões) Todos os resultados indicam que a clonagem e a caracterização da HSP83 do Trypanosoma cruzi foram alcançadas.

Abstract (Objective) Trypanosoma cruzi, a causal agent of Chagas’ disease, is a parasite whose life cycle alternates between an invertebrate (triatomine) and a vertebrate (mammal) host. Various studies have shown that in T. cruzi, HSP83 (a homolog of HSP90) is essential for cell division and control of the response to thermal stress. This investigation focused on studying the cloning, bioinformatic characterization, and expression of the heat shock protein HSP83 T. cruzi gene for further cell signaling studies. (Methodology) RNA was extracted from T. cruzi epimastigotes (EPm6 clone, MHOM/VE/2007/ 6c) using a commercial kit. The cDNA encoding HSP83 was determined using RT-PCR on extracted mRNA, for which the primers were designed based on the HSP83 sequence of T. cruzi strain CL Brener. Cloning was performed using pGEM®T-Easy and subcloned into the expression vector pQE30. Sequence and bioinformatic characterization were performed. The gene was expressed, and the recombinant protein was purified using affinity chromatography and identified through immunoblotting. (Results) Sequence analysis showed similarity to the gene encoding HSP83 from Trypanosoma cruzi, and HSP domains and B epitopes in the sequence were also observed. After 3 hours of induction with IPTG, a recombinant protein with an approximate weight of 83 kDa was obtained. The immunoblotting reaction with hyperimmune anti-T. cruzi epimastigote serum helped detect a single band with a molecular weight of nearly 83 kDa. (Conclusions) All results indicate that the cloning and characterization of HSP83 from Trypanosoma cruzi was achieved.

Abstract Shrimp aquaculture has rapidly expanded in the last decades, representing an important economic activity worldwide; however, infections (bacterial and viral) are one of the biggest challenges in aquaculture activity. Acute Hepatopancreatic Necrosis Disease (AHPND) is a bacterial disease that affects shrimp farm ponds and occurs during the first 30 days after stocking in shrimp ponds, causing mortalities of up to 70 - 100 %. Microbiota has an important role in shrimp aquaculture and disease control. In the present study, pirA and pirB genes were amplified by PCR to confirm AHPND and non-AHPND in shrimp hepatopancreas; samples were sequenced using the MiSeq platform targeting the V3-V4 16S ribosomal (rRNA) hypervariable regions. Results demonstrated a diverse microbiota in the non-AHPND group, mainly composed of phyla Proteobacteria, Firmicutes, and Actinobacteria. Furthermore, Proteobacteria was the dominant phyla in the hepatopancreas infected with AHPND, while Firmicutes tended to decrease. Vibrio was the most abundant at the genus level, prevailing in some genera like Corynebacterium, Weissella, Lactobacillus, Photobacterium, and Geobacillus. Results suggest that AHPND causes dysbiosis in the hepatopancreatic microbiota, and the Firmicutes phylum could require to be restored under such scenario.

Resumen La producción acuícola de camarón se ha expandido rápidamente en las últimas décadas, convirtiéndose en una importante actividad económica a nivel mundial. Sin embargo, algunas infecciones bacterianas y virales son un gran desafío para esta actividad. La Necrosis Hepatopancreática Aguda (AHPND) es una enfermedad bacteriana que afecta a los estanques de granjas camaroneras y se presenta durante los primeros 30 días después de la siembra en estanques de camarones, causando una mortalidad del 70 - 100 %. La microbiota tiene un papel importante en la producción del camarón y el control de enfermedades. En el presente estudio, los genes pirA y pirB fueron amplificados por PCR para confirmar tanto organismos positivos como negativos a AHPND en hepatopáncreas de camarones. Posteriormente se secuenciaron las regiones hipervariables ribosómicas (ARNr) V3-V4 del gen 16S utilizando la plataforma MiSeq. Los resultados demostraron una microbiota diversa en el grupo negativo a AHPND, compuesto principalmente por filos Proteobacteria, Firmicutes y Actinobacteria. Además, Proteobacteria fue el filo dominante en el hepatopáncreas infectado con AHPND, mientras que Firmicutes tendió a disminuir. Vibrio fue el género más abundante, prevaleciendo sobre algunos géneros como Corynebacterium, Weissella, Lactobacillus, Photobacterium y Geobacillus. Los resultados sugieren que la enfermedad de AHPND causa disbiosis en la microbiota hepatopancreática, y el filo Firmicutes podría requerir ser restaurado en dicho escenario.

Abstract Ovine Enzootic Abortion (OEA) is caused by Chlamydia abortus and has recently been recognized as endemic in Mexico. Due to the impact of OEA on intensive lamb production, it is necessary to establish a sanitary control plan based on an efficient diagnosis. The loop-mediated isothermal amplification technique (LAMP) is a molecular test that can specifically identify C. abortus. The aim of this study was to standardize the LAMP technique for the specific detection of C. abortus in ovine vaginal exudates. Primers were identified and modified from an electronic database. The effective conditions of the technique were determined using synthetic DNA and positive biological samples by PCR as references. The assay detection limit was 1 × 10-5 of genetic material, equivalent to 8.5 copies. The standardized LAMP test is proposed as a novel molecular tool for the rapid and specific diagnosis of C. abortus infection from ovine vaginal exudate samples in Mexico.

Resumen El Aborto Enzoótico Ovino (AEO) es causado por Chlamydia abortus y ha sido reconocido recientemente como endémico en México. Debido al impacto del AEO en la producción intensiva de corderos, es necesario establecer un plan de control sanitario basado en un diagnóstico eficiente. La técnica de amplificación isotérmica mediada por bucle (LAMP) es una prueba molecular que puede identificar específicamente a C. abortus. El objetivo de este estudio fue estandarizar la técnica de LAMP para la detección específica de C. abortus a partir de exudado vaginal ovino. Los cebadores fueron identificados y modificados a partir de una base de datos electrónica. Las condiciones efectivas de la técnica se determinaron con ADN sintético y de muestras biológicas positivas por PCR como referencia. El límite de detección de la prueba fue de 1 × 10-5 de material genético, equivalente a 8.5 copias. Se propone el test de LAMP estandarizado como una nueva herramienta molecular para el diagnóstico rápido y específico de la infección por C. abortus a partir de muestras de exudado vaginal ovino en México.

SUMMARY: Cerebral hemorrhage (CH) presents a significant risk for cognitive decline. Our research aimed to assess the impact of Apricot Kernel Oil (AKO) on cognitive function and inflammatory response in a hemorrhagic stroke model using Wistar rats, and to examine their correlation. Fifty Wistar rats were randomly assigned to five groups: a control group, an AKO supplemented group (receiving 400 mg/kg AKO), a CH group (with induced CH), and CH groups supplemented with 200 and 400 mg/kg AKO, each containing 10 rats. Cognitive function was evaluated using the Y-electric maze test across these groups. Additionally, enzyme-linked immunosorbent assay and real-time PCR were utilized to analyze the protein and gene expression profiles of inflammatory and apoptotic factors in the brain. Compared to the control group, rats in the CH group took longer to master the maze criteria (P<0.05). Moreover, the CH group exhibited significantly increased levels of pro-inflammatory cytokines interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α, and decreased levels of anti-inflammatory cytokines IL-4 and IL-10 compared to the control group (P<0.05). Conversely, rats in the CH+400 mg/kg AKO group showed opposite trends in electrical simulation time, inflammatory factor protein, and gene expression profile (Bcl- 2, BAX, Caspase-3, and p53) compared to the CH group (P<0.05). Our findings suggest that AKO extract may ameliorate cognitive dysfunction following CH in rats, potentially through modulation of inflammatory factors secreted by microglia.

RESUMEN: La hemorragia cerebral (HC) presenta un riesgo significativo de deterioro cognitivo. Nuestra investigación tuvo como objetivo evaluar el impacto del aceite de semilla de albaricoque (ASA) en la función cognitiva y la respuesta inflamatoria en un modelo de accidente cerebrovascular hemorrágico utilizando ratas Wistar, y examinar su correlación. Se asignaron aleatoriamente cincuenta ratas Wistar a cinco grupos: un grupo control, un grupo suplementado con ASA (que recibió 400 mg/kg de ASA), un grupo HC (con HC inducido) y grupos HC suplementados con 200 y 400 mg/kg de ASA, cada uno. La función cognitiva se evaluó mediante la prueba del laberinto eléctrico Y en estos grupos. Además, se utilizaron un ensayo inmunoabsorbente ligado a enzimas y una PCR en tiempo real para analizar los perfiles de expresión de proteínas y genes de factores inflamatorios y apoptóticos en el cerebro. En comparación con el grupo de control, las ratas del grupo HC tardaron más en dominar los criterios del laberinto (P<0,05). Además, el grupo HC exhibió niveles significativamente mayores de citocinas proinflamatorias interleucina-1β (IL-1β), IL-6 y factor de necrosis tumoral- α, y niveles reducidos de citocinas antiinflamatorias IL-4 e IL-10 en comparación al grupo control (P<0,05). Por el contrario, las ratas del grupo HC+400 mg/kg ASA mostraron tendencias opuestas en el tiempo de simulación eléctrica, la proteína del factor inflamatorio y el perfil de expresión génica (Bcl-2, BAX, Caspasa-3 y p53) en comparación con el grupo HC (P <0,05). Nuestros hallazgos sugieren que el extracto de ASA puede mejorar la disfunción cognitiva después de la HC en ratas, potencialmente mediante la modulación de factores inflamatorios secretados por la microglia.

Abstract: The main objective of this study is to characterize the virus associated with faba bean (Vicia faba L.) crops in the Department of Nariño (Colombia). Faba bean is an important food source, but it faces challenges from viral diseases such as "pod mottle." This disease manifests with symptoms such as leaf chlorosis, dwarfism, and pod lesions. Viral characterization is conducted through seed transmission tests, mechanical transmission, RT-PCR, and transmission electron microscopy (TEM). The results show that the virus morphology consists of spherical and enveloped particles that range between 80-100nm in diameter. RT-PCR and sequencing of the nucleocápside gen (558bp) reveals the virus is Orthotospovirus tomatomaculae (Tomato spotted wilt virus) with a 97% homology. Seed transmission is 2% while mechanical transmission is 40%. This is the first report of an Orthotospovirus affecting faba bean in Nariño, posing a risk to food security.

Resumen: El principal objetivo de este estudio es caracterizar el virus asociado al cultivo de haba (Vicia faba L.) en el Departamento de Nariño (Colombia). El cultivo de haba es una fuente importante de alimento, pero enfrenta desafíos por enfermedades virales como la "viruela de la vaina". Esta enfermedad se manifiesta con síntomas como clorosis en las hojas, enanismo y lesiones en la vaina. La caracterización del virus se realiza mediante pruebas de transmisión por semilla, transmisión mecánica, RT-PCR y microscopía electrónica de transmisión (TEM). Los resultados muestran que la morfología del virus consiste en partículas esféricas y envueltas de entre 80-100nm en diámetro. RT-PCR y secuenciación del gen nucleocápside (558pb) revela que el virus es Orthotospovirus tomatomaculae (Tomato spotted wilt virus) con una homología de 97%. La transmisión por semilla es de 2% y mecánicamente del 40%. Este es el primer informe de un Orthotospovirus afectando el haba en Nariño, lo que representa un riesgo para la seguridad alimentaria.

Abstract Background Merkel cell polyomavirus (MCPyV), a human polyomavirus that is unequivocally linked to merkel cell carcinoma (MCC), has been found in association with keratinocytes carcinomas (KC), especially basal cell carcinoma (BCC) and cutaneous squamous cell carcinoma (cSCC). Nevertheless, there is scarce information about the possible involvement of MCPyV in the development of KC. Objectives To assess the presence of MCPyV DNA and Large-T Antigen (LT-Ag) via Polymerase Chain Reaction (PCR) and Immunohistochemistry (IHC) in cases of KC, and to correlate its presence with immunohistochemical markers p16, p53, and ki67, tumor type and subtype, sun-exposed location, and epidemiological data. Methods The prevalence of MCPyV DNA, LT-Ag, and immunohistochemical markers p16, p53, and ki67 was assessed by PCR and Immunohistochemistry (IHC) in 127 cases of KC, these results were correlated with tumor type and subtype, sun-exposed location, and epidemiological data. Results The MCPyV DNA was detected in 42.57% (43 of 101) cases by PCR, the LT-Ag was detected in 16.4% (20 of 122) of cases, p16 in 81.5% (97 of 119), p53 in 66.4% (83 of 125), ki67 in 89% (73 of 82). No correlation between MCPyV LT-Ag and DNA confronted with tumor type, subtype, location site, and immunohistochemical markers was found. A single correlation between the MCPyV LT-Ag and cSCC tumors and peri-tumoral lymphocyte cells was noted. Study limitations Further steps need to be taken to better evaluate the MCPyV influence and its possible role in KC carcinogenesis, as the evaluation of the virus genome state, the gene sequence that encodes LT-Ag in the KC tumor cells, and in situ hybridization for viral DNA or RNA in these cells. Conclusions Despite the frequent detection of MCPyV in KC, the data available so far does not support the hypothesis of a causal relationship between them. MCPyV, , (MCPyV) MCC, MCC (MCC) (KC) BCC (BCC cSCC. . (cSCC) Nevertheless LargeT Large T LTAg LT Ag (LT-Ag (PCR IHC (IHC p ki subtype sunexposed sun exposed LTAg, Ag, ki6 12 4257 42 57 42.57 43 (4 101 164 16 4 16.4 20 (2 122 p1 815 81 5 81.5 97 (9 119, 119 119) p5 664 66 66.4 83 (8 125, 125 125) 89 73 (7 82. 82 82) site peritumoral peri tumoral noted carcinogenesis state them (MCPyV (MCC (KC (cSCC 1 425 42.5 ( 10 16. 2 8 81. 9 11 6 66. 7 42.

SUMMARY OBJECTIVE: Coronary artery disease is the leading cause of death worldwide. Vascular endothelial growth factor is known to induce endothelial cell migration and proliferation, increase vascular permeability, and modulate thrombogenicity. The aim of this study is to investigate the relationship between the VEGF insertion/deletion (I/D) variant (rs35569394) and coronary artery disease susceptibility in the Turkish population. METHODS: A total of 206 subjects, including 106 coronary artery disease patients and 100 controls, were included in this study. The VEGF I/D variant was genotyped using the polymerase chain reaction method. RESULTS: The frequency of the I/I, I/D, and D/D genotypes was 35.84 versus 37%, 33.97 versus 36%, and 30.19 versus 27% in patients and the control group, respectively. VEGF I/D genotype and allele distribution were not statistically significant between coronary artery disease patients and controls (p>0.05). There was no significant difference between VEGF I/D genotype distribution and patient characteristics including age, gender, disease duration, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglyceride, history of hypertension, history of diabetes mellitus, and smoking (p>0.05). CONCLUSION: This study suggests that the VEGF I/D variant is not a predisposing factor to coronary artery disease disease in a Turkish sample. OBJECTIVE worldwide proliferation permeability thrombogenicity insertiondeletion insertion deletion ID I D (I/D rs35569394 rs (rs35569394 population METHODS 20 subjects 10 method RESULTS II I/I DD 3584 35 84 35.8 37 37% 3397 33 97 33.9 36 36% 3019 30 19 30.1 27 group respectively p>0.05. p005 p p>0.05 . 0 05 (p>0.05) age gender duration highdensity high density cholesterol lowdensity low triglyceride hypertension mellitus CONCLUSION sample rs3556939 (rs3556939 2 1 358 3 8 35. 339 9 33. 301 30. p00 p>0.0 (p>0.05 rs355693 (rs355693 p0 p>0. (p>0.0 rs35569 (rs35569 p>0 (p>0. rs3556 (rs3556 p> (p>0 rs355 (rs355 (p> rs35 (rs35 (p rs3 (rs3 (rs

Resumen El presente es un estudio retrospectivo, observacional, cuantitativo y descriptivo. Se evaluó la utilidad de la proteína C reactiva (PCR), la procalcitonina (PCT) y la relación PCR/PCT como marcadores de riesgo de sepsis, sumados al aclaramiento a las 72 h, como pronóstico de mortalidad y permanencia en unidades de cuidados intensivos (UCI). Se incluyeron 23 pacientes. Se clasificaron según qSOFA y se elaboraron curvas ROC. Se obtuvo un área bajo la curva de 0,79 para PCT. El valor umbral de PCT>0,88 ng/mL predice riesgo de sepsis con 77,78% de sensibilidad y 83,33% de especificidad. Utilizando PCR<31,23 mg/dL se obtuvo como parámetros destacados un 88,89% de sensibilidad y 83,33% de valor predictivo negativo, sin diferencias significativas (Mann-Whitney p<0,05) entre los grupos de sobrevivientes y óbitos y estadía prolongada vs. no prolongada. Se postula PCR como screening y PCT como marcador de riesgo de sepsis.

Resumo Este é um estudo retrospectivo, observacional, quantitativo e descritivo. Foi avaliada a utilidade da proteína C reativa (PCR), da procalcitonina (PCT) e da relação PCR/PCT como marcadores de risco de sepse junto com o clareamento em 72 horas, como preditores de mortalidade e permanência em unidades de terapia intensiva (UTI). Vinte e três pacientes foram incluídos. Eles foram classificados de acordo com o qSOFA e foram elaboradas curvas ROC, destacando uma área sob a curva de 0,79 para PCT. O valor limite de PCT>0,88 ng/mL prediz o risco de sepse com sensibilidade de 77,78% e especificidade de 83,33%. Utilizando PCR>31,23 mg/dL, obtiveram-se como parâmetros em destaque 88,89% de sensibilidade e 83,33% de valor preditivo negativo. Não houve diferenças significativas (Mann-Whitney p<0,05) entre os grupos de sobreviventes e óbitos e permanência prolongada vs. não prolongada. A PCR é postulada como triagem e a PCT como marcador de risco de sepse.

Abstract This is a retrospective, observational, quantitative and descriptive study. The utility of C reactive protein (CRP), procalcitonin (PCT) and the CRP/PCT ratio as sepsis risk markers was evaluated and these, added to clearance at 72 hours, as predictors of mortality and permanence in intensive care units (ICU). Twenty-three patients were included. They were classified according to qSOFA, and ROC curves were prepared, highlighting an area under the curve of 0.79 for PCT. The threshold value of PCT>0.88 ng/mL predicts the of sepsis with 77.78% sensitivity and 83.33% specificity. Using CRP>31.23 mg/dL, 88.89% sensitivity and 83.33% negative predictive value were obtained as outstanding parameters. No significant differences (Mann-Whitney p<0.05) were found between survivors and dead and prolonged vs. non-prolonged stay groups. CRP is postulated for screening and PCT as a sepsis risk marker.

ABSTRACT This study aimed to determine the coexistence and food sources of adult mosquitoes (Diptera: Culicidae) in a rural health center in Piura, Peru by using a descriptive cross-sectional design. Entomological techniques were used to capture and identify mosquitoes, and molecular biotechnology techniques were used to identify food sources. A total of 793 specimens of the Culex and Aedes genera were found coexisting, 789 (99.5%) were Culex quinquefasciatus, 607 (76.9%) were males and 182 (23.1%) were females. Likewise, 4 (100%) corresponded to Aedes aegypti females. The food sources of Aedes aegypti were Homo sapiens sapiens, and Homo sapiens sapiens and Canis familiaris were the food sources of Culex quinquefasciatus. This study provides evidence that rural health centers could be acting as foci of arbovirosis, with the risk that people who come for different ailments could contract diseases transmitted by C. quinquefasciatus and A. aegypti.

RESUMEN Con el objetivo del estudio fue determinar la coexistencia y fuentes de alimentación de mosquitos adultos (Diptera: Culicidae) en un centro de salud rural de Piura en Perú, se realizó un estudio descriptivo transversal. Se usaron técnicas entomológicas para capturar e identificar mosquitos, y técnicas de biotecnología molecular para identificar las fuentes de alimentación. Un total de 793 ejemplares de los géneros Culex y Aedes se encontraron coexistiendo, 789 (99,5%) corresponden a Culex quinquefasciatus, 607 (76,9%) fueron machos y 182 (23,1%) hembras. Así mismo, 4 (100%) corresponden a Aedes aegypti hembras. Las fuentes de alimentación de Aedes aegypti fueron Homo sapiens sapiens, y de Culex quinquefasciatus fueron Homo sapiens sapiens y Canis familiaris. Este estudio proporciona evidencia de que los centros de salud rurales estarían actuando como focos de arbovirosis, existiendo el riesgo de que personas que acuden por distintas dolencias, puedan contraer enfermedades transmitidas por C. quinquefasciatus y A. aegypti.

ABSTRACT Objective. To determine the feeding behavior of Aedes aegypti in dengue outbreaks in two rural areas of Peru during the Yaku cyclone and El Niño phenomenon of 2023. Material and methods. Eight blood samples (8 pools) were obtained from the abdomen of 80 Aedes aegypti specimens captured in the rural districts of Querecotillo and Marcavelica during the Yaku cyclone and El Niño dengue outbreaks. DNA was extracted from the analyzed samples, then a PCR was directed at the CytB gene as a genetic marker and the PCR products were enzymatically digested with the restrictases Hae III and Mwo I. The PCR-RFLP products were visualized by agarose gel electrophoresis at 4%. Results. DNA was obtained from all samples and a 358 bp amplicon was obtained as a PCR product. Likewise, the only RFLP found in Hae III was from Homo sapiens sapiens (233 and 125 bp). RFLP was not found in Hae III of Gallus gallus and RFLP in Mwo I of Canis familiaris and Mus musculus. Conclusion. Aedes aegypti showed conserved anthropophilic feeding behavior in dengue outbreaks in rural areas during the Yaku cyclone and El Niño.

RESUMEN Objetivos. Determinar la alimentación del Aedes aegypti en brotes de dengue de dos zonas rurales del Perú durante el ciclón Yaku y El Niño Global del 2023. Material y métodos. Se analizaron ocho muestras de sangre (8 pooles) obtenidas del abdomen de 80 especímenes Aedes aegypti capturados en los distritos rurales de Querecotillo y Marcavelica durante brotes de dengue acontecidos en el ciclón Yaku y en El Niño Global. Se extrajo ADN de las muestras analizadas, se llevó a cabo una PCR dirigida al gen CytB como marcador genético y los productos PCR fueron digeridos enzimáticamente con las restrictasas Hae III y Mwo I. Los productos PCR-RFLP fueron visualizados por electroforesis en gel de agarosa al 4%. Resultados. Se obtuvo ADN de todas las muestras y como producto PCR un amplicón de 358 pb. Así mismo, el único RFLP en Hae III observado fue el de Homo sapiens sapiens (233 y 125 pb). No se observó RFLP en Hae III de Gallus gallus y RFLP en Mwo I de Canis familiaris y Mus musculus. Conclusión. En brotes de dengue de zonas rurales, durante el ciclón Yaku y en El Niño Global, el Aedes aegypti presentó un comportamiento alimenticio antropofílico conservado.

Resumen La mastitis bovina es una enfermedad que afecta a las ganaderías de pequeños y medianos productores de los cantones Cayambe y Pedro Moncayo, Provincia de Pichincha-Ecuador. El tratamiento de esta enfermedad resulta complicado debido a la variedad de microorganismos que la provocan. El presente estudio se enfocó en la determinación molecular por medio de reacción en cadena de la polimerasa (PCR) de los agentes etiológicos de la mastitis. Esta técnica presenta múltiples ventajas al reconocer familia, género y especie de microorganismos, y es un método capaz de detectar genes de resistencia de antibióticos, lo que resulta importante al momento de diagnosticar y tratar enfermedades. El objetivo de esta investigación se centró en la identificación de bacterias causantes de la mastitis bovina, utilizando pruebas bioquímicas y moleculares. Las pruebas bioquímicas como tinción Gram, Catalasa, Coagulasa, y Agar Manitol Sal fueron eficientes para obtener cepas puras y determinar el género de algunas bacterias. Se utilizaron primers específicos (RNA16S) para la identificación molecular de 9 agentes etiológicos causantes de la enfermedad en las unidades productivas. Los microorganismos encontrados fueron; Staphylococcus pasteuri, Staphylococcus warneri, Staphylococcus sp., Staphylococcus epidermidis, Staphylococcus aureus, Staphylococcus saprophyticus, Sphingomonas sp., Streptococcus dysgalactiae, Streptococcus uberis, la mayoría presentes en mastitis clínica. Para detectar genes de resistencia se utilizaron primers específicos, de los cuales 7 muestras presentaron el gen para resistencia a blaTEM (β-lactámicos) y 6 muestras presentaron el gen para resistencia a tetA (tetraciclinas). Se identificó multirresistencia en las especies: Staphylococcus pasteuri, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus uberis, Sphingomonas sp.

Abstract Bovine mastitis is a disease that affects the farms of small and medium producers in the cantons of Cayambe and Pedro Moncayo, Pichincha Province-Ecuador. Treating this disease is not easy due to the different microorganisms that cause it. This study focused on the molecular determination by means of polymerase chain reaction (PCR) of the etiological agents of mastitis, having multiple advantages when recognizing family, gender and species of microorganisms. It is a method capable of detecting resistance genes of antibiotics, an important analysis when diagnosing and treating diseases. The aim of this research is to identify bacteria causing bovine mastitis by using biochemical and molecular tests. Biochemical tests such as: Gram staining, Catalase, Coagulase, and Mannitol Salt Agar were efficient to obtain pure strains and determine the gender of some bacteria. Specific primers (RNA16S) were used for the molecular identification of 9 etiological agents causing the disease in the productive units. The microorganisms found were Staphylococcus pasteuri, Staphylococcus warneri, Staphylococcus sp., Staphylococcus epidermidis, Staphylococcus aureus, Staphylococcus saprophyticus, Sphingomonas sp., Streptococcus dysgalactiae, Streptococcus uberis, mostly present in clinic mastitis. To detect resistance genes, specific primers were used, of which 7 samples presented the gene for resistance to blaTEM (β-lactam) and 6 samples presented the gene for resistance to tetA (tetracyclines). Multi-resistance was identified in the species Staphylococcus pasteuri, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus uberis, Sphingomonas sp.

Resumen El virus dengue es un virus endémico en Argentina que, si bien inicialmente se consideró que no era neu rotrópico, actualmente se reconoce que tiene neuroinva sión, condicionando así una prevalencia de manifesta ciones neurológicas de hasta el 15% entre los enfermos. Aun siendo considerados síntomas de gravedad, existe subdiagnóstico de encefalitis por dengue debido a su variada forma de presentación clínica. Las manifesta ciones neurológicas de la encefalitis por dengue pueden abarcar desde fiebre y cefalea hasta alteraciones del nivel de conciencia y convulsiones. Si bien el líquido cefalorraquídeo (LCR) puede hallarse normal en hasta un tercio de los casos, lo habitual es que presente aumento de concentración de proteínas y pleocitosis. En cuanto a los métodos de neuroimagen, los hallazgos suelen ser variados e inespecíficos, e incluso pueden ser normales hasta en 40-50% de los casos. Se presentan 3 casos de encefalitis por dengue diag nosticados en un hospital universitario de Buenos Ai res, Argentina, en donde la presentación clínica varió desde desorientación témporo-espacial hasta estatus convulsivo refractario con diferentes presentaciones en el LCR pero todos con PCR positivo para dengue y con neuroimágenes sin alteraciones.

Abstract Dengue virus is an endemic virus in Argentina that, although it was initially considered to be non-neuro tropic, it is currently recognized to be neuroinvasive; thus conditioning a prevalence of neurological mani festations of up to 15% among patients. Even being considered severe symptoms, there is underdiagnoses of dengue encephalitis due to its varied clinical presenta tion. Neurological manifestations of dengue encephalitis can range from fever and headache to altered levels of consciousness and seizures. Although the cerebrospinal fluid may be normal in up to a third of cases, it usually presents increased protein concentration and pleocyto sis. Regarding neuroimaging methods, the findings are usually varied and nonspecific, and can even be normal in up to 40-50% of cases. We present three cases of dengue encephalitis di agnosed in a university hospital in Buenos Aires, Ar gentina, where the clinical presentation varied from temporal-spatial disorientation to refractory convulsive status with different presentations in the cerebrospinal fluid but all with positive PCR for dengue in it and with normal neuroimaging.